Tjie Kok The Remediation Abstrak 2010
ICECS 2010
2010 3rd International Conference on
17-19，October，2010
Kunming, China
i
2010 3rd International Conference on
Environmental and Computer Science
(ICECS)
Copyright and Reprint Permission: Abstracting is permitted with credit to the source. Libraries are permitted to
photocopy beyond the limit of U.S. copyright law for private use of patrons those articles in this volume that
carry a code at the bottom of the first page, provided the per-copy fee indicated in the code is paid through
Copyright Clearance Center, 222 Rosewood Drive, Danvers, MA 01923. For other copying, reprint or
republication permission, write to IEEE Copyrights Manager, IEEE Operations Center, 445 Hoes Lane, P.O. Box
1331, Piscataway, NJ 08855-1331.
Copyright ©2010 by the Institute of Electrical and Electronics Engineers. All rights reserved.
© 2010 IEEE. Personal use of this material is permitted. However, permission to reprint/republish this material
for advertising or promotional purposes or for creating new collective works for resale or redistribution to
servers or lists, or to reuse any copyrighted component of this work in other works must be obtained from the
IEEE.
IEEE Catalog Number: CFP1075I-PRT
ISBN: 978-1-4244-7630-5
IEEE Catalog Number: CFP1075I-ART
ISBN: 978-1-4244-7632-9
IEEE Catalog Number: CFP1075I-CDR
ISBN: 978-1-4244-7631-2
Publisher: Institute of Electrical and Electronics Engineers, Inc.
Printed in Chengdu, China
ii
PREFACE
Dear Distinguished Delegates and Guests,
The Organizing Committee warmly welcomes our distinguished delegates and guests to the 2010 3rd
International Conference on Environmental and Computer Science (ICECS 2010) held on October 17-19, 2010
in Kunming, China.
ICECS 2010 is sponsored by Asia-Pacific Chemical, Biological & Environmental Engineering Society (APCBEES)
and International Association of Computer Science and Information Technology (IACSIT), and supported by
APCBEES, IACSIT Members and scholars from universities all round the world. If you have attended a
conference sponsored by APCBEES and IACSIT before, you are aware that the conferences together report the
results of research efforts in a broad range of chemical, biological and environmental engineering society.
These conferences are aimed at discussing with all of you the wide range of problems encountered in present
and future high technologies. ICECS 2010 is organized to gather members of our international community
scientists so that researchers from around the world can present their leading-edge work, expanding our
community’s knowledge and insight into the significant challenges currently being addressed in that research.
The conference Program Committee is itself quite diverse and truly international, with membership from the
Americas, Europe, Asia, Africa and Oceania.
This proceeding records the fully refereed papers presented at the conference. The main conference themes
and tracks are Environmental and Computer Engineering. The main goal of these events is to provide
international scientific forums for exchange of new ideas in a number of fields that interact in-depth through
discussions with their peers from around the world. Both inward research; core areas of Environmental and
Computer Engineering and outward research; multi-disciplinary, inter-disciplinary, and applications will be
covered during these events.
The conference has solicited and gathered technical research submissions related to all aspects of major
conference themes and tracks. All the submitted papers in the proceeding have been peer reviewed by the
reviewers drawn from the scientific committee, external reviewers and editorial board depending on the
subject matter of the paper. Reviewing and initial selection were undertaken electronically. After the rigorous
peer-review process, the submitted papers were selected on the basis of originality, significance, and clarity
for the purpose of the conference. The selected papers and additional late-breaking contributions to be
presented as lectures will make an exiting technical program. The conference program is extremely rich,
featuring high-impact presentations.
The high quality of the program – guaranteed by the presence of an unparalleled number of internationally
recognized top experts – can be assessed when reading the contents of the program. The conference will
therefore be a unique event, where attendees will be able to appreciate the latest results in their field of
expertise, and to acquire additional knowledge in other fields. The program has been structured to favor
interactions among attendees coming from many diverse horizons, scientifically, geographically, from
academia and from industry. Included in this will to favor interactions are social events at prestigious sites.
We would like to thank the program chairs, organization staff, and the members of the program committees
for their work. Thanks also go to Editor Ms. Feng Ying, International Association of Computer Science and
Information Technology, for her wonderful editorial service to this proceeding.
We are grateful to all those who have contributed to the success of ICECS 2010. We hope that all participants
and other interested readers benefit scientifically from the proceedings and also find it stimulating in the
process. Finally, we would like to wish you success in your technical presentations and social networking.
iii
We hope you have a unique, rewarding and enjoyable week at ICECS 2010 in Kunming, China.
With our warmest regards,
The Organizing Committees
October 17-19, 2010
Kunming, China.
iv
Organizing Committees
Conference Chairs
Prof. Safaa S. Mahmoud, Ain Shams University, Egypt
Prof. Jong Soo Cho, Catholic University of Korea, Korea
Prof. Yucong Duan, University of Bourgogne, France
Program Committee Chairs
Rita Sandip Dhodapkar, Natonal Environmental Engineering Research Institute, India
Farhad Nejadkoorki, Yazd University, Iran
Luo Zhenfei, University of Electronics Science and Technology of China, China
Yin Zhixi, North University of China, China
Prasad Girijesh, University of Ulster, UK
Hu Jiankun, RMIT University, Australia
Conference Steering Committee
Liu Yuhua, Huazhong Normal University, China
Kung, Chih-hsien, Chang-Jung Christian University, Taiwan
Ir. David B.L. Bong, Universiti Malaysia Sarawak, Malaysia
Wang Hongjun, Southwest Jiaotong University, China
G.S.Tomar, Vikrant Institute of Technology and Management, India
Publication Chair
Li Kai, Guizhou University, China
Secretariat Contacts
ICECS 2010 Website: http://www.icecs.org
ICECS 2010 E-mail: icecs@vip.163.com
xiii
Reviewers
Technical Committee
Prof. Smita Rajpal, Institute of Technology and Management, India
Prof. Wang Dianhui, La Trobe University, Australia
Prof. Shobha G, R.V College of Enginering, India
Prof. Tartaglia Angelo, Information Engineering Faculty, Politecnico di Torino, Italy
Prof. Nazir Ahmad Zafar, University of Central Punjab, Pakistan
Prof. Yang Laurence T., St. Francis Xavier University, Canada
Prof. Jun Guo, East China Normal University, China
Prof. Matsui Masayoshi, National Instisute of Advanced Industrial Science and Technology, Japan
Prof. Nasro Min Allah, COMSATS Institute of Information Technology, Pakistan
Prof. Nashat Mansour, Lebanese American University, Lebanon
Prof. Jun-qing Li, Liaocheng University, China
Prof. Assassa Ghazy, King Saud University, Kingdom of Saudia Arabia
Prof. Chih-ming Kung, Shih Chien University, Taiwan
Prof. Wei Guo, Tianjin University, China
Prof. Amir Masoud Rahmani, Islamic Azad University, Iran
Prof. Somkait Udomhunsakul, King Mongkutt’s Institute of Technology Ladkrabang, Thailand
Prof. Debojyoti Mitra, Sir Padampat Singhania University, India
Muhammad Jamil Khan, Gomal University, Pakistan
Raghu Reddy Yedduladoddi, Institute of Technology Rochester, USA
Adel Smeda, Accounting University of Al-Jabel, Libya
Khairuddin Hashim, Tenaga Nasional University, Malaysia
Thanakorn Sornkaew, Rangsit University, Thailand
Oliver Moravcik, Slovak University of Technology, Slovakia
G K Viju, Karary University, Sudan
S.Britto R.Kumar, Bishop Heber College(Autonomous), India
Yuhanis Yusof, Universiti Utara Malaysia, Malaysia
Rui Gomes, Rui Manuel da Silva Gomes, Turkey
xiv
Contents
Preface
Organizing Commitees
iii
xii
ICECS 2010 Session 1
The Remediation of Copper Ions by Shoot Cultures of Solanum Melongena and Its Influence on The
Production of Phytosterols and Phytosteroids
1
Tjie Kok
Domestic Wastewater Sludge as a Major Substrate for Cellulase Production by Two Different Fungal
Strains
6
Md. Zahangir Alam, Afifah Drani and Suleyman A. Muyibi
Development of Land Suitability Model for Irrigation Management (LSM) Using Remote Sensing, GIS,
GPS and Field Studies: A Case study of a Part of Nagarjunasagar Command Area, Andhra Pradesh,
INDIA
9
Rajasekhar Reddy.N
On-Line Monitoring and Control of a Partially Packed BAF System for Nitrogen Removal in a Synthetic
Wastewater
14
Pramanik BIPLOB, Suja FATIHAH and Zain SHAHAROM
18
New Approaches in Urban Air Quality Monitoring
Farhad Nejadkoorki
Performance Analysis of MAC for Wireless Body Area Sensor Networks
21
Sai Anand Gopalan and Jong-Tae Park
Adsorption of Bovine Serum Albumin on nano-Hydroxyapatite
26
Jie Lian, Wenhua Song, Liying Li and Jinhua Zhang
Adsorption of Bovine Serum Album on Mesoporous Silica MCM-41
29
Jinhua Zhang, Wenhua Song, Liying Li and Jie Lian
Optimizing Process Water Use at Two Large Australian Manufacturing Companies
33
B.A. Agana, J.D Orbell and D. Reeve
A Construction Approach for PIM of Multimodal Web User Interfaces
39
Yun Shi and Hebiao Yang
ICECS 2010 Session 2
Regional Eco-Environmental Information Service System Based on Open Source Projects
43
LI Guangyu, ZHOU Kefa, SUN Li, WANG Jinlin and WANG Qianfeng
Application of TOPSIS Model Based on AHP Weight to Beiyun River Ecosystem Health Assessment
49
Wenxian Guo and Hongxiang Wang
A Field Study Using Iodine Tracer to Characterize Chemical Contaminants Penetrated into Groundwater
from Vadose Zone
v
54
2010 3rd International Conference on Environmental and Computer Science (ICECS 2010)
The Remediation of Copper Ions by Shoot Cultures of Solanum Melongena and Its
Influence on The Production of Phytosterols and Phytosteroids
Tjie Kok
Faculty of Biotechnology, Mathematics and Science Department,
University of Surabaya, Surabaya, Indonesia
+ Corresponding author. Tel.: + 6281 75088277; fax: +6231 2981278
E-mail address: tjie_kok@yahoo.com
Abstract— This study aimed to observe the remediation of
copper ions by shoot cultures of Solanum melongena and its
influence on the production of
phytosterol(s) and
phytosteroid(s). The cultures were able to remove 46.6-66.5%
of the ions from media containing 80-320 µM of the ions and
accumulated them in their biomass. Application of copper ions
concentration higher than that of control Murashige and
Skoog media modified with the addition of benzyladenine 4
µg/L caused a decrease in total free sterols content of the
cultures. The higher the copper ions concentration, the higher
the decrease in total free sterols content was. Spotting the
hydrolysate extracts of the cultures on tin layer
chromatography (TLC) plate, developing using a mixture
of CHCl3:CH3OH (10:1), and spraying the spots by
Dragendorff’s reagent−followed by heating for 5 minutes at
1000C, there was a blue spot with Rf (retardation factor) value
similar to that of standard (solasodine). The chromatograms of
gas chromatograph (GC) showed that the retention time (Rt)
of certain substance contained in the extracts is similar to that
of solasodine. Verification using mass spectrometer (MS)
showed that the ionic fragments of the substance are similar to
those of standard solasodine (dilute). It could then be predicted
that the shoot cultures of Solanum melongena were be able to
produce solasodine. Further verification is needed to ensure
whether it was solasodine or not; it can be conducted by using
nuclear magnetic resonance (NMR).
Keywords-Solanum melongena, copper ion, phytosterols,
phytosteroids, shoot culture.
I.
INTRODUCTION
Plant cell cultures, as well as its intact plant can give
certain responses on the presence of certain metal ions in the
medium. Cultures of Chlorella vulgaris could remove 75%
of 0.5 ppm copper ions from the medium [1]. Hairy root
cultures of Armoracia rusticana were able to remove
62.4% of the copper ions from initial value of 20.5 ppm
copper ions [2]. Copper tolerant callus cultures of Nicotiana
tabacum regenerated plants that were able to grow in the
presence of 6.0 ppm copper ions [3].
Free sterols content of shoot cultures of Costus
speciosus increased in media containing up to 20 ppm
copper ions, the highest was at 5 ppm of the ions (120%),
compared to the control (0.006 ppm). In media containing 15 ppm copper ions, the content of bound sterols increased up
to 119-213%, whilst at concentration 20 ppm the ions
978-1-4244-7631-2/10/$26.00 C 2010 IEEE
caused a decrease in that content (85%). Adding up to 20
ppm copper ions into the media caused an increase of total
(sum of free and bound) sterols content of that cultures, the
highest was at 5 ppm of that ions (134%). The cultures
remediated 26-38% copper ions from media containing 1-20
ppm of the ions [4].
Callus cultures of Agave amaniensis produced squalene,
sterols and some sapogenin steroids, where hecogenin is the
major constituent [5]. Copper ions at concentration of 0.6
ppm in the media caused an increase of total sapogenin
steroids content in callus cultures of Agave amaniensis [5].
Using a relatively high concentration of calcium (2.7-4.5
mM), magnesium (4.3-5.3 mM), cobalt (0.4-1.0 µM) and
copper (7.2-12.3 µM) ions simultaneously in media,
inhibition of the sapogenin steroids production in callus
cultures of Agave amaniensis was observed [6].
Application of 2.5 ppm copper ions to callus cultures of
Costus speciosus was able to increase the content of free
and bound sterols; addition of 1 ppm and 20 ppm copper
ions into media decreased the free
sterols content; at concentration 5, 10 and 20 ppm copper
ions caused a decrease in the content of bound sterols [7].
Callus cultures of Solanum mammosum produced some
sterols (cholesterol, sitosterol, stigmasterol, campesterol),
but solasodine was not produced by that cultures [8].
Coppers ions are one of the essential micronutrients for
normal plant growth. Excess of copper ions, however,
causes a range of deleterious influences including inhibition
of photosynthesis, pigment synthesis and damage to the
plasma membrane permeability and other metabolic
disturbances [3].
II.
EXPERIMENTAL
A. Materials
The shoot cultures of Solanum melongena were
cultivated on modified Murashige and Skoog media with the
addition of 4 µg/L benzyladenine. Cultures were maintained
in the light using Philips TL40W at 250C. Subculturing of
the stock-shoots was performed every 4 weeks [5].
In order to study the accumulation of copper ions and its
influence on the production of phytosterols and
phytosteroids, the cultures were transferred to a series of 20
ml media containing different concentration of copper ions
and cultivated for 4 weeks under the same conditions before
V4-1
2010 3rd International Conference on
17-19，October，2010
Kunming, China
i
2010 3rd International Conference on
Environmental and Computer Science
(ICECS)
Copyright and Reprint Permission: Abstracting is permitted with credit to the source. Libraries are permitted to
photocopy beyond the limit of U.S. copyright law for private use of patrons those articles in this volume that
carry a code at the bottom of the first page, provided the per-copy fee indicated in the code is paid through
Copyright Clearance Center, 222 Rosewood Drive, Danvers, MA 01923. For other copying, reprint or
republication permission, write to IEEE Copyrights Manager, IEEE Operations Center, 445 Hoes Lane, P.O. Box
1331, Piscataway, NJ 08855-1331.
Copyright ©2010 by the Institute of Electrical and Electronics Engineers. All rights reserved.
© 2010 IEEE. Personal use of this material is permitted. However, permission to reprint/republish this material
for advertising or promotional purposes or for creating new collective works for resale or redistribution to
servers or lists, or to reuse any copyrighted component of this work in other works must be obtained from the
IEEE.
IEEE Catalog Number: CFP1075I-PRT
ISBN: 978-1-4244-7630-5
IEEE Catalog Number: CFP1075I-ART
ISBN: 978-1-4244-7632-9
IEEE Catalog Number: CFP1075I-CDR
ISBN: 978-1-4244-7631-2
Publisher: Institute of Electrical and Electronics Engineers, Inc.
Printed in Chengdu, China
ii
PREFACE
Dear Distinguished Delegates and Guests,
The Organizing Committee warmly welcomes our distinguished delegates and guests to the 2010 3rd
International Conference on Environmental and Computer Science (ICECS 2010) held on October 17-19, 2010
in Kunming, China.
ICECS 2010 is sponsored by Asia-Pacific Chemical, Biological & Environmental Engineering Society (APCBEES)
and International Association of Computer Science and Information Technology (IACSIT), and supported by
APCBEES, IACSIT Members and scholars from universities all round the world. If you have attended a
conference sponsored by APCBEES and IACSIT before, you are aware that the conferences together report the
results of research efforts in a broad range of chemical, biological and environmental engineering society.
These conferences are aimed at discussing with all of you the wide range of problems encountered in present
and future high technologies. ICECS 2010 is organized to gather members of our international community
scientists so that researchers from around the world can present their leading-edge work, expanding our
community’s knowledge and insight into the significant challenges currently being addressed in that research.
The conference Program Committee is itself quite diverse and truly international, with membership from the
Americas, Europe, Asia, Africa and Oceania.
This proceeding records the fully refereed papers presented at the conference. The main conference themes
and tracks are Environmental and Computer Engineering. The main goal of these events is to provide
international scientific forums for exchange of new ideas in a number of fields that interact in-depth through
discussions with their peers from around the world. Both inward research; core areas of Environmental and
Computer Engineering and outward research; multi-disciplinary, inter-disciplinary, and applications will be
covered during these events.
The conference has solicited and gathered technical research submissions related to all aspects of major
conference themes and tracks. All the submitted papers in the proceeding have been peer reviewed by the
reviewers drawn from the scientific committee, external reviewers and editorial board depending on the
subject matter of the paper. Reviewing and initial selection were undertaken electronically. After the rigorous
peer-review process, the submitted papers were selected on the basis of originality, significance, and clarity
for the purpose of the conference. The selected papers and additional late-breaking contributions to be
presented as lectures will make an exiting technical program. The conference program is extremely rich,
featuring high-impact presentations.
The high quality of the program – guaranteed by the presence of an unparalleled number of internationally
recognized top experts – can be assessed when reading the contents of the program. The conference will
therefore be a unique event, where attendees will be able to appreciate the latest results in their field of
expertise, and to acquire additional knowledge in other fields. The program has been structured to favor
interactions among attendees coming from many diverse horizons, scientifically, geographically, from
academia and from industry. Included in this will to favor interactions are social events at prestigious sites.
We would like to thank the program chairs, organization staff, and the members of the program committees
for their work. Thanks also go to Editor Ms. Feng Ying, International Association of Computer Science and
Information Technology, for her wonderful editorial service to this proceeding.
We are grateful to all those who have contributed to the success of ICECS 2010. We hope that all participants
and other interested readers benefit scientifically from the proceedings and also find it stimulating in the
process. Finally, we would like to wish you success in your technical presentations and social networking.
iii
We hope you have a unique, rewarding and enjoyable week at ICECS 2010 in Kunming, China.
With our warmest regards,
The Organizing Committees
October 17-19, 2010
Kunming, China.
iv
Organizing Committees
Conference Chairs
Prof. Safaa S. Mahmoud, Ain Shams University, Egypt
Prof. Jong Soo Cho, Catholic University of Korea, Korea
Prof. Yucong Duan, University of Bourgogne, France
Program Committee Chairs
Rita Sandip Dhodapkar, Natonal Environmental Engineering Research Institute, India
Farhad Nejadkoorki, Yazd University, Iran
Luo Zhenfei, University of Electronics Science and Technology of China, China
Yin Zhixi, North University of China, China
Prasad Girijesh, University of Ulster, UK
Hu Jiankun, RMIT University, Australia
Conference Steering Committee
Liu Yuhua, Huazhong Normal University, China
Kung, Chih-hsien, Chang-Jung Christian University, Taiwan
Ir. David B.L. Bong, Universiti Malaysia Sarawak, Malaysia
Wang Hongjun, Southwest Jiaotong University, China
G.S.Tomar, Vikrant Institute of Technology and Management, India
Publication Chair
Li Kai, Guizhou University, China
Secretariat Contacts
ICECS 2010 Website: http://www.icecs.org
ICECS 2010 E-mail: icecs@vip.163.com
xiii
Reviewers
Technical Committee
Prof. Smita Rajpal, Institute of Technology and Management, India
Prof. Wang Dianhui, La Trobe University, Australia
Prof. Shobha G, R.V College of Enginering, India
Prof. Tartaglia Angelo, Information Engineering Faculty, Politecnico di Torino, Italy
Prof. Nazir Ahmad Zafar, University of Central Punjab, Pakistan
Prof. Yang Laurence T., St. Francis Xavier University, Canada
Prof. Jun Guo, East China Normal University, China
Prof. Matsui Masayoshi, National Instisute of Advanced Industrial Science and Technology, Japan
Prof. Nasro Min Allah, COMSATS Institute of Information Technology, Pakistan
Prof. Nashat Mansour, Lebanese American University, Lebanon
Prof. Jun-qing Li, Liaocheng University, China
Prof. Assassa Ghazy, King Saud University, Kingdom of Saudia Arabia
Prof. Chih-ming Kung, Shih Chien University, Taiwan
Prof. Wei Guo, Tianjin University, China
Prof. Amir Masoud Rahmani, Islamic Azad University, Iran
Prof. Somkait Udomhunsakul, King Mongkutt’s Institute of Technology Ladkrabang, Thailand
Prof. Debojyoti Mitra, Sir Padampat Singhania University, India
Muhammad Jamil Khan, Gomal University, Pakistan
Raghu Reddy Yedduladoddi, Institute of Technology Rochester, USA
Adel Smeda, Accounting University of Al-Jabel, Libya
Khairuddin Hashim, Tenaga Nasional University, Malaysia
Thanakorn Sornkaew, Rangsit University, Thailand
Oliver Moravcik, Slovak University of Technology, Slovakia
G K Viju, Karary University, Sudan
S.Britto R.Kumar, Bishop Heber College(Autonomous), India
Yuhanis Yusof, Universiti Utara Malaysia, Malaysia
Rui Gomes, Rui Manuel da Silva Gomes, Turkey
xiv
Contents
Preface
Organizing Commitees
iii
xii
ICECS 2010 Session 1
The Remediation of Copper Ions by Shoot Cultures of Solanum Melongena and Its Influence on The
Production of Phytosterols and Phytosteroids
1
Tjie Kok
Domestic Wastewater Sludge as a Major Substrate for Cellulase Production by Two Different Fungal
Strains
6
Md. Zahangir Alam, Afifah Drani and Suleyman A. Muyibi
Development of Land Suitability Model for Irrigation Management (LSM) Using Remote Sensing, GIS,
GPS and Field Studies: A Case study of a Part of Nagarjunasagar Command Area, Andhra Pradesh,
INDIA
9
Rajasekhar Reddy.N
On-Line Monitoring and Control of a Partially Packed BAF System for Nitrogen Removal in a Synthetic
Wastewater
14
Pramanik BIPLOB, Suja FATIHAH and Zain SHAHAROM
18
New Approaches in Urban Air Quality Monitoring
Farhad Nejadkoorki
Performance Analysis of MAC for Wireless Body Area Sensor Networks
21
Sai Anand Gopalan and Jong-Tae Park
Adsorption of Bovine Serum Albumin on nano-Hydroxyapatite
26
Jie Lian, Wenhua Song, Liying Li and Jinhua Zhang
Adsorption of Bovine Serum Album on Mesoporous Silica MCM-41
29
Jinhua Zhang, Wenhua Song, Liying Li and Jie Lian
Optimizing Process Water Use at Two Large Australian Manufacturing Companies
33
B.A. Agana, J.D Orbell and D. Reeve
A Construction Approach for PIM of Multimodal Web User Interfaces
39
Yun Shi and Hebiao Yang
ICECS 2010 Session 2
Regional Eco-Environmental Information Service System Based on Open Source Projects
43
LI Guangyu, ZHOU Kefa, SUN Li, WANG Jinlin and WANG Qianfeng
Application of TOPSIS Model Based on AHP Weight to Beiyun River Ecosystem Health Assessment
49
Wenxian Guo and Hongxiang Wang
A Field Study Using Iodine Tracer to Characterize Chemical Contaminants Penetrated into Groundwater
from Vadose Zone
v
54
2010 3rd International Conference on Environmental and Computer Science (ICECS 2010)
The Remediation of Copper Ions by Shoot Cultures of Solanum Melongena and Its
Influence on The Production of Phytosterols and Phytosteroids
Tjie Kok
Faculty of Biotechnology, Mathematics and Science Department,
University of Surabaya, Surabaya, Indonesia
+ Corresponding author. Tel.: + 6281 75088277; fax: +6231 2981278
E-mail address: tjie_kok@yahoo.com
Abstract— This study aimed to observe the remediation of
copper ions by shoot cultures of Solanum melongena and its
influence on the production of
phytosterol(s) and
phytosteroid(s). The cultures were able to remove 46.6-66.5%
of the ions from media containing 80-320 µM of the ions and
accumulated them in their biomass. Application of copper ions
concentration higher than that of control Murashige and
Skoog media modified with the addition of benzyladenine 4
µg/L caused a decrease in total free sterols content of the
cultures. The higher the copper ions concentration, the higher
the decrease in total free sterols content was. Spotting the
hydrolysate extracts of the cultures on tin layer
chromatography (TLC) plate, developing using a mixture
of CHCl3:CH3OH (10:1), and spraying the spots by
Dragendorff’s reagent−followed by heating for 5 minutes at
1000C, there was a blue spot with Rf (retardation factor) value
similar to that of standard (solasodine). The chromatograms of
gas chromatograph (GC) showed that the retention time (Rt)
of certain substance contained in the extracts is similar to that
of solasodine. Verification using mass spectrometer (MS)
showed that the ionic fragments of the substance are similar to
those of standard solasodine (dilute). It could then be predicted
that the shoot cultures of Solanum melongena were be able to
produce solasodine. Further verification is needed to ensure
whether it was solasodine or not; it can be conducted by using
nuclear magnetic resonance (NMR).
Keywords-Solanum melongena, copper ion, phytosterols,
phytosteroids, shoot culture.
I.
INTRODUCTION
Plant cell cultures, as well as its intact plant can give
certain responses on the presence of certain metal ions in the
medium. Cultures of Chlorella vulgaris could remove 75%
of 0.5 ppm copper ions from the medium [1]. Hairy root
cultures of Armoracia rusticana were able to remove
62.4% of the copper ions from initial value of 20.5 ppm
copper ions [2]. Copper tolerant callus cultures of Nicotiana
tabacum regenerated plants that were able to grow in the
presence of 6.0 ppm copper ions [3].
Free sterols content of shoot cultures of Costus
speciosus increased in media containing up to 20 ppm
copper ions, the highest was at 5 ppm of the ions (120%),
compared to the control (0.006 ppm). In media containing 15 ppm copper ions, the content of bound sterols increased up
to 119-213%, whilst at concentration 20 ppm the ions
978-1-4244-7631-2/10/$26.00 C 2010 IEEE
caused a decrease in that content (85%). Adding up to 20
ppm copper ions into the media caused an increase of total
(sum of free and bound) sterols content of that cultures, the
highest was at 5 ppm of that ions (134%). The cultures
remediated 26-38% copper ions from media containing 1-20
ppm of the ions [4].
Callus cultures of Agave amaniensis produced squalene,
sterols and some sapogenin steroids, where hecogenin is the
major constituent [5]. Copper ions at concentration of 0.6
ppm in the media caused an increase of total sapogenin
steroids content in callus cultures of Agave amaniensis [5].
Using a relatively high concentration of calcium (2.7-4.5
mM), magnesium (4.3-5.3 mM), cobalt (0.4-1.0 µM) and
copper (7.2-12.3 µM) ions simultaneously in media,
inhibition of the sapogenin steroids production in callus
cultures of Agave amaniensis was observed [6].
Application of 2.5 ppm copper ions to callus cultures of
Costus speciosus was able to increase the content of free
and bound sterols; addition of 1 ppm and 20 ppm copper
ions into media decreased the free
sterols content; at concentration 5, 10 and 20 ppm copper
ions caused a decrease in the content of bound sterols [7].
Callus cultures of Solanum mammosum produced some
sterols (cholesterol, sitosterol, stigmasterol, campesterol),
but solasodine was not produced by that cultures [8].
Coppers ions are one of the essential micronutrients for
normal plant growth. Excess of copper ions, however,
causes a range of deleterious influences including inhibition
of photosynthesis, pigment synthesis and damage to the
plasma membrane permeability and other metabolic
disturbances [3].
II.
EXPERIMENTAL
A. Materials
The shoot cultures of Solanum melongena were
cultivated on modified Murashige and Skoog media with the
addition of 4 µg/L benzyladenine. Cultures were maintained
in the light using Philips TL40W at 250C. Subculturing of
the stock-shoots was performed every 4 weeks [5].
In order to study the accumulation of copper ions and its
influence on the production of phytosterols and
phytosteroids, the cultures were transferred to a series of 20
ml media containing different concentration of copper ions
and cultivated for 4 weeks under the same conditions before
V4-1